Hepatocurative effect of Emblica officinalis against CYP 450 bio-activation hepatotoxicity of drugs

ABSTRACT

The present invention relates to a composition useful for hepatocurative effect against CYP 450 bio-activation hepatotoxicity induced by drugs, said composition comprising an extract from Emblica officinalis and optionally pharmaceutically acceptable additives and method of treating drug induced hepatotoxicity.

CROSS REFERENCE TO RELATED APPLICATION

This application is a division of Ser. No. 10/106,119, filed Mar. 27,2002, and which is incorporated in its entirety herein by reference.

FIELD OF THE PRESENT INVENTION

The present invention relates to a composition useful for hepatocurativeeffect against CYP 450 bio-activation hepatotoxicity induced by drugs,said composition comprising an extract from Emblica officinalis andoptionally pharmaceutically acceptable additives and method of treatingdrug induced hepatotoxicity.

BACKGROUND OF THE PRESENT INVENTION

Liver disorders are still the major health hazards both in urban andrural areas of the world. Despite scientific advances in ourunderstanding of hepatotoxicity, and leads provided by traditionalsystem of medicine, we do not have yet any effective entities to cureliver derangement more importantly those which are caused by a varietyof drugs. The Indian Council of Medical Research, New Delhi in itsrevised research programme on traditional medicines, has adopted liverdiseases as one among six thrust areas for multidisciplinary study.

The disorders of the liver may be classified into acute or chronichepatitis (inflammatory liver diseases), hepatosis (non-inflammatorydisorders). The acute condition is often followed by liver cirrhosis aswell as hepatic coma with grave prognosis. Liver cirrhosis as suchaccounts amongst the ten top fatal diseases in the world. Exposure ofhumans to a variety of agents such as chemicals and drugs (xenobiotics),many natural compounds, viral and bacterial pathogens with attendantpredisposable conditions, etc. are considered responsible for hepaticinsufficiency.

There are large group of drugs which on repeated administration produceliver toxicity. These are mediated primarily by bioactivation so thatthe products of parent drug are toxic. Another class of drugs inducetoxicity by causing membrane rupture or DNA damage and by interferingwith protein synthesis. One of the important categories of drugs are theanti-TB drugs which when taken regularly cause hepatotoxicity.

Tuberculosis is prevelent in all counteries of the world--tropical,subtropical and colder regions. The chemotherapy of tuberculosis isimportant and challenging, because the disease is often chronic and thetoxicity due to anti-TB drugs pose therapeutic problems. The disordersof the liver caused during the treatment of tuberculosis, by knownantimicrobial agents range from jaundice to the fibrosis of the liver.Several cases progress to the chronic form of disease or have afulminant course and prove fatal. Three drugs i.e., rifampicin,pyrazinamide and isoniazid comprise first choice treatment oftuberculosis. These are to be administered for long period of time andproduce liver dysfunction leading to toxicity.

Jaundice is amongst the most prominent incidence of their adversereactions. The characteristic pathology is the bridging and multilobularnecrosis. Hypersensitivity to these drugs leads to hepatitis. Multidrugtreatment also poses special problems. Rifampicin causes liver damage.Disturbances in liver function is more if it is combined with isoniazid.Pyrazinamide is the most toxic of the three anti-TB drugs. Continuationof the drugs in combination after symptoms of hepatic dysfunction haveappeared tends to increase further the severity of damage. Severehepatic injury leading to death has been reported in patients receivingthese drugs (Slivka, I L, Farmakol Toksikol-1989: 52;82-85)

In our traditional system of medicines (Ayurveda), use of severalmedicinal plants have been prescribed for alleviating liver disorders.There are nearly forty indigenous polyherbal formulations from more than100 plants enjoying reputation of being hepatoprotectives. However, nonehave been specified as a therapeutic agent, which is able to protect theliver from injury due to treatment of anti-TB drugs. This owes partly tothe fact that reports are scanty with regard to evaluation ofplants/plant products, which would seem focussed specially againsthepatic injury caused by drugs which produce toxicity as a result ofbioactivation.

There is thus a growing interest in the development of herbal entitiesconsidered relatively safe for alleviating liver disorders specificallycaused by the anti-TB drugs.

Emblica officinalis Gaertn. (Hindi: Amla) (Euphorbiaceae)is widespreadin India, Ceylon, Malaya and China. The tree is common in mixeddeciduous forests of India ascending to 4500 ft on the hills, cultivatedin gardens and homeyards. It is a small or medium sized deciduous tree,fruits depressed globose, ½ to 1 inch in diameter, fleshy, and containssix trigonows seeds. The fruit is sour and is occasionally eaten raw.The fruit pulp contains (%); moisture 81.2, protein 0.5, fat 0.1,mineral matter 0.7, Ca 0.005, Phosphorus 0.02 and Iron 1.2 mg/100 gm,nicotinic acid 0.2 mg/100 gm, vitamin C 600 mg/100 mg. (Medicinal plantsof India, Satyavati et al (ed.), ICMR, new Delhi, 1976, p 377). Thepotent vitamin C-like activity has been located in the low molecularweight hydrolysable tannins. Four such compounds emblicanin-A,emblicanin-B, punigluconin and pedunculagin have been isolated from thefresh pericarp. The first two compounds are naturally occurringgalloellagi-tannins (Ghosal, et al, IndJChem, 1996:353:941-948;Bhattacharya et al, Phytomedicine, 2000: 7: 173-175)

The fruit is acrid, cooling, and diuretic. Dried fruit is useful inhaemorrhage, diarrhoea and dysentry. It has been extensively use inanemia, liver diseases and dyspepsea. A fermented liquor prepared fromthe fruits is used in jaundice. Fruits are a reputed Ayurvedic rasayan(revitaliser, biological response modifier) (Sharma P. V. Dravyagunavijnana, Chaukhamba Sanskrit Sansthan, Varanasi, 1978). Severalpharmacological properties are also reported. Leaf extracts have beenfound to be anti-inflammatory (Summanen et al, Planta Medica, 1993:59:666), antioxidant (Jose and Kuttan, Clin. Biochem. Nutr, 1995:19:63-70),hypolipidemic (Mathw:eta.backslash.,JEthnopharmacol, 1996:50 :61-68),cell growth inhibition (Psatima et al, ACS Symp. Ser, 1998, p701).Hepatoprotective activity of Emblica officinalis extracts against achemical viz., carbon tetrachoride induced liver toxicity has beendemonstrated (Jose J K & Kutten R, J. Ethnopharmacology 2000:72; 135-40;Bhattacharya et al, Phytomerdicine 2000:7:173-5).

The article by Sharma et al (Hum Exp. toxicol 2000:19; 337-84) suggeststhat Emblica Officinalis prevents genotoxicity induced by benzopyrines.Benzopyrene is one of the prominent environmental carcinogen which is aspecific substrate for CYP 450 1A1. Both are clastogenic. However, theliver toxicity produced by the drugs including anti-TB drugs isdependent on a great measure to their bio-activation through multipleCYP isoforms, the most prominent being CYP450 3A4. There are severalagents, which reduce CYP levels or reverse the micronuclei formation butare not hepatoprotective. To cite an example, applicants have developeda molecule, piperine which is a specific inhibitor of CYP 450 1A1, butis not hepatoprotective. Similarly, there are several known compounds,which reverse the genotoxicity but are not hepatoprotective. Thereforethe decrease in CYP 1A1 or reversal of clastrogencity can not beconstrued as hepatoprotection. For example, jaundice (hepato-biliarydysfunction) has not been correlated with genotoxicity, rather it may bean early event in the onset of liver toxicity. In the present article, acasual relationship between CYP decrease or genotoxicity has not beenrelated to attenuation of clinical pathology usually seen in symptoms ofhepatotoxicity.

In the present invention, the Applicants provide protection againsthepatotoxicity produced by all such drugs, which are bio-activated bymultiple CYP isoforms as indicated by clinical parameters inserum/liver. Besides, we claim that the clinical parameters showingtoxicity are reversed even if the symptoms of genotoxicity may not beginto appear. For example decrease in abnormal rise of serum Bilirubin,which may be attributed to a protective effect also due to othercellular factors such as membrane stabilization, as revealed in primarymonolayer cultures of liver cells.

Thus, we claim preparations from Emblica officinalis which are superiorso far as their systemic effects are manifested in clinical profile(serum/liver parameters) which correlate to their hepatoprotectiveprofile.

OBJECT OF THE PRESENT INVENTION

The main object of the present invention is to develop a hepatocurativecomposition against CYP 450 bio-activation hepatotoxicity induced bydrugs.

Another main object of the present invention is to develop ahepatocurative composition against CYP 450 bio-activation hepatotoxicityinduced by anti-TB drugs.

Yet another object of the present invention is to develop a compositionfrom fruit Emblica officinalis for hepatocurative effect.

Still another object of the present invention is to develop acomposition from fruit Emblica officinalis for hepatocurative effectagainst CYP 450 bio-activation hepatotoxicity.

Still another object of the present invention is to develop a method ofpreparing an extract from fruit Emblica officinalis.

Still another object of the present invention is to develop a method fortreating a subject for CYP 450 and free radical mediated hepatotoxicitycaused by drugs using composition comprising an extract from Emblicaofficinalis.

Still another object of the present invention is to develop a method ofusing hepatocytes to understand the effect of extract from fruit Emblicaofficinalis.

SUMMARY OF THE INVENTION

The present invention relates to a composition useful for hepatocurativeeffect against CYP 450 bio-activation hepatotoxicity induced by drugs,said composition comprising an extract from Emblica officinalis andoptionally pharmaceutically acceptable additives and method of treatingdrug induced hepatotoxicity.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

Accordingly, the present invention relates to a composition useful forhepatocurative effect against CYP 450 bio-activation hepatotoxicityinduced by drugs, said composition comprising an extract from Emblicaofficinalis and optionally pharmaceutically acceptable additives andmethod of treating drug induced hepatotoxicity.

In one embodiment of the present invention, a composition useful forhepatocurative effect against CYP 450 bio-activation hepatotoxicityinduced by drugs, said composition comprising an extract from Emblicaofficinalis and optionally pharmaceutically acceptable additives.

In another embodiment of the present invention, wherein said additivesare selected from a group of nutrients comprising proteins,carbohydrates, sugar, talc, magnesium stearate, cellulose, calciumcarbonate, starch-gelatin paste, and/or pharmaceutically acceptablecarrier, excipient, diluent, or solvent.

In yet another embodiment of the present invention, wherein saidcomposition is administered orally.

In still another embodiment of the present invention, wherein saidextract and additives are in the ratio ranging between 1:1 to 1:10.

In still another embodiment of the present invention, wherein saidadditives have no effect on the hepatocurative effect of the saidextract.

In still another embodiment of the present invention, wherein saidextract is prepared in a solvent selected from a group comprisingaqueous, aqueous-ethanolic, ethanolic, ketonic, ethereal, halogenatedsolvents.

In still another embodiment of the present invention, wherein saidcomposition shows tanin content in the range of 8.5 to 15%.

In still another embodiment of the present invention, wherein, saidcomposition for the oral route is in form of capsule, tablet, syrup,concentrate, powder, granule, aerosol, and/or beads.

In further another embodiment of the present invention, a method ofpreparing composition comprising an extract from Emblica officinalis andoptionally pharmaceutically acceptable additives, said method comprisingsteps of adding polar solvent to the fruit Emblica Officinalis to obtainthe extract and optionally adding pharmaceutically acceptable additives.

In another embodiment of the present invention, wherein said fruit isincubated with polar solvent at room temperature for about 15-25 hours.

In yet another embodiment of the present invention, wherein said extractis from fresh and/or semi dried fruits of Emblica Officinalis.

In still another embodiment of the present invention, wherein saidextract and additives are in the ratio ranging between 1:1 to 1:10.

In still another embodiment of the present invention, wherein saidextract is prepared in a solvent selected from a group comprisingaqueous, aqueous-ethanolic, ethanolic, ketonic, ethereal, halogenatedsolvents.

In still another embodiment of the present invention, wherein, saidcomposition for the oral route is in form of capsule, tablet, syrup,concentrate, powder, granule, aerosol, and/or beads.

In further embodiment of the present invention, a method of treating asubject for CYP 450 and free radical mediated hepatotoxicity caused bydrugs using composition comprising an extract from Emblica officinalisand optionally pharmaceutically acceptable additives.

In another embodiment of the present invention, introducing drugtoxicity in hepatocytes.

In yet another embodiment of the present invention, adding saidcomposition to said hepatocytes exposed to drug hepatotoxicity.

In still another embodiment of the present invention, measuring changesin the level of liver/serum markers to estimate hepatocurative effect ofthe said composition.

In still another embodiment of the present invention, wherein saidmethod is particularly effective against hepatotoxicity caused byanti-TB drugs.

In still another embodiment of the present invention, wherein saidcomposition is not effective against hepatotoxicity which is independentof bio-activation by CYP 450.

In still another embodiment of the present invention, wherein saidcomposition is administered orally.

In still another embodiment of the present invention, wherein, saidcomposition for the oral route is in form of capsule, tablet, syrup,concentrate, powder, granule, aerosol, and/or beads.

In still another embodiment of the present invention, wherein saidcomposition is useful for treating animals or human beings.

In still another embodiment of the present invention, wherein saidcomposition has no adverse effect on health.

In still another embodiment of the present invention, wherein said drugsare selected from a group comprising Paracetamol, CCI.sub.4, and anti-TBdrugs.

In still another embodiment of the present invention, wherein saidanti-TB drugs are selected from a group comprising Rifampicin,Pyrazinamide, and isoniazid.

In still another embodiment of the present invention, wherein saidcomposition controls abnormal rise in the clinical pathological symptomsrevealed by serum/liver markers serving as indices of hepatic damagebesides control of high levels of Bilirubin.

In still another embodiment of the present invention, wherein saidmethod uses hepatocyte culture for ideal insight.

In still another embodiment of the present invention, wherein said drugsis used at cytotoxic levels to produce valid and reproducible results inliver cells.

In still another embodiment of the present invention, wherein saidcomposition is useful for treating animals or human beings.

In still another embodiment of the present invention, wherein saidcomposition has no adverse effect on health.

In still another embodiment of the present invention, wherein saidcomposition shows restoration of hepatocyte viability.

In still another embodiment of the present invention, wherein saidmethod shows prevention of cell membrane leakage.

In still another embodiment of the present invention, wherein saidcomposition shows about 96% hepatocurative effect against combinedeffect of anti-TB drugs of Rifampicin, and Isoniazid.

In still another embodiment of the present invention, wherein saidcomposition reverses the leakage of glutamate pyruvate transaminase(GPT) from hepatocyte.

In still another embodiment of the present invention, wherein saidcomposition shows hepatocurative effect of about 96% against combinedeffect of anti-TB drugs of Rifampicin, isoniazid, and pyrazinamide.

In still another embodiment of the present invention, wherein saidcomposition shows about 94% hepatocurative effect against rise in lipidPeroxidation (LPO) induced by combination of anti-TB drugs Rifampicin,isoniazid, and pyrazinamide.

In still another embodiment of the present invention, wherein saidcomposition shows about 96% decrease of serum Bilirubin as ahepatocurative effect against combination of anti-TB drugs Rifampicin,isoniazid, and pyrazinamide.

In still another embodiment of the present invention, wherein saidmethod helps restore liver function to normal.

In still another embodiment of the present invention, wherein dosage ofsaid composition is ranging between 50-250 mg/kg.

In still another embodiment of the present invention, wherein saidmethod is used for hepatocurative effect against drugs causing liverdysfunction, including anti-TB drugs.

In further embodiment of the present invention, the applicants provideprotection against hepatotoxicity produced by all drugs, which arebio-activated by multiple CYP isoforms as indicated by clinicalparameters in serum/liver. Besides, we claim that the clinicalparameters showing toxicity are reversed even if the symptoms ofgenotoxicity may not begin to appear. For example decrease in abnormalrise of serum Bilirubin, which may be attributed to a protective effectalso due to other cellular factors such as membrane stabilization, asrevealed in primary monolayer cultures of liver cells.

Further, applicants have made use of their expertise and years toresearch to establish that Emblica Officinalis (Alma) cureshepatotoxicity induced by drugs that is restricted to CYP 450bio-activation hepatotoxicity.

Thus, applicants claim preparations from Emblica officinalis which aresuperior so far as their systemic effects are manifested in clinicalprofile (serun/liver parameters) which correlate to theirhepatoprotective profile.

In further embodiment of the present invention, it relates topreparations and methods of preparation and use of such products whichrestores the normal liver function against drug induced toxicity as aresult of bio-activation of drugs applicable with particular relevanceto anti-TB drug(s) induced liver toxicity. The compositions and methodsof the present invention increase biological defence mechanism of thetissue, improve recovery from dysfunctional states of the liver afterprolonged challenge of anti-TB drugs.

In another embodiment of the present invention, the compositions andmethods of the present invention contain one of the extracts/fractionsof Emblica officinalis fruit as an essential ingredient. Theseextracts/fractions may be obtained from fresh or semi dried fruits ofEmblica officinalis. The compositions are formulated with more than oneextracts and combined in any weight ratios. The preferred weight ratiosinclude 1:1,1:2,1:1:1:, 1:2:2.2:1:2:, 2:2:1.

In still another embodiment of the present invention, it is related topreparation and use of products from Emblica officinalis, which restoresnormal liver function against drug induced toxicity caused as a resultof bio-activation of drugs applicable with particular relevance toanti-TB drug(s) induced liver toxicity. The products of the inventioncomprise aqueous, aqueous-ethanolic, ethanolic, ketonic, ethereal,halogenated solvents extracts/fractions from Emblica officinalis,obtained either from fresh or semi-dried fruits. These contain 8.5-15%of tannin content. It also relates to preparation of compositions ofsuch products in different proportions of more than one ingredient.These products either alone or in combination are intended to be usedagainst drug induced liver toxicity as represented by specific mechanismunderlying liver disorders and usually manifested in clinical conditionsof liver dysfunction.

In still another embodiment of the present invention, the preparationseither alone or in composition are also intended to be used againstanti-TB drug(s).The use of such products as developed in the presentinvention controls the abnormal rise in clinical pathological symptomsrevealed by serum/liver markers serving as indices of hepatic damagebesides control of high levels of Bilirubin.

In still another embodiment of the present invention, in the developmentof the present invention the ample information has been utilized whichexists regarding the advances in our understanding of mechanismsresponsible for hepatotoxic injury due to drugs. More importantly thechoice of a suitable model for the evaluation of anti-toxic profile ofany substance is also crucial.

In still another embodiment of the present invention, a large body ofinformation has been gained in favor of the present invention by usingliver cell (hepatocyte) cultures, which ideally provide an insight intothe mechanism of a toxin-induced impairment of hepato-biliarydysfunction because this model allows use of a test compounds (such asanti-TB drugs) to be used at cytotoxic level so that a valid andreproducible toxicity is generated. The in vitro cell culture model isof significant interest in ascertaining the mechanisms of toxicity andits reversal by protective agents.

In still another embodiment of the present invention, liver cells areconsidered as system of choice which have found ample application in theevaluation of cyto-cum geno-toxicity of chemicals and drugs (Nakagawaand Tayama, Arch Toxicol, 1995:69:208) and as such have been used in theevaluation of hepatoprotective profiles of the present invention. Themechanisms are revealed in critical biochemical functions of liver cellswhich are sensitive indicators of drug (s) toxicity. (Tomasi et al,Toxicol/Vtf/zo/: 15:178-183). Both cellular lysis (measured by leakageof transaminases enzymes and lactated dehydrogenase from the cells) andthe metabolic competence of the tissue are modified as a function ofboth the duration and concentration of the drugs (Goethals et al FundmAppl Toxicol 1984:4:441-450).

In still another embodiment of the present invention, the preparations(alone or in combination) act in a specific manner. These act againsttoxicity produced by drugs including anti-TB drugs which requirebioactivation by hepatic cytochrome P 450 dependent mixed functionoxidases. Cytochrome P450 have been shown to be involved in the livertoxicity (Anundi I, Lindros K O, Pharmacol Toxicol 1992; 70;453-458).Participation of CYP 450 dependent oxidation of drugs includinganti-tubercular drugs rifampicin, isoniazid, pyrazinamide in liver isreported (Ono et al, Biol Pharm Bull 1998:21:421-425).

In still another embodiment of the present invention, that thepreparations mentioned in the present invention act in a specific manneris further evidenced by the observation that these preparations have notbeen found effective against galactosamine induced toxicity which is notdependent upon the intervention of mixed function oxidases. Of seriousconcern is the toxicity produced by use of some drugs in combinationsuch as anti-tubercular drugs. Preparations alone or in combination,prevent not only (a) rifampicin (b) isoniazid (c) pyrazinamide inducedtoxicity but also various combinations of these such as (a)rifampicin+isoniazid (b) rifampicin+pyrazinamide (c)isoniazid+pyrazinamide and (d) rifampicin+isoniazid+pyrazinamidetoxicity. The metabolic activation of drugs including anti-TB drugsalone or in combination is also accompanied by reactive intermediateswhich may be free radical/active metabolites/free oxy radicals through avariety of cellular oxidative metabolic pathways. An alteredoxidative/antioxidative profile is closely associated with production ofdrug (s) induced hepatic injury (Sodhi et al, Hum Exp Toxicol 1997;16;315-321). The efficacy of the products of the present invention isfurther shown by their anti-lipoperoxidant (anti-oxidant) profile. Byusing the preparations of the present invention a decrease in theaccumulation of excess levels of the product of oxygen metabolism hasbeen revealed.

In still another embodiment of the present invention, also included arethe preventing role of the products developed herein, against cellmembrane leakage and restoration of cell viability in challenged livertissues caused as a result of toxic menifestations. Preparations act ina specific manner in as much as they prevent toxicity produced bybioactivation of drug (s) and combination of drugs (s) as described inthe above art. The products described have no cytotoxicity and on theother hand enhance overall biological defense systems per se.

Although the invention has been described in conjunction with specificembodiments, it is evident that many alternatives and variations will beapparent to those skilled in the art in light of the foregoingdescription. Accordingly, the invention is intended to embrace all ofthe alternatives and variations that fall within the spirit and scope ofthe appended claims. The foregoing references are hereby incorporated byreference.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

FIG. 1 shows liver toxicity wherein plant extract attenuates Rifampicininduced hepatotoxicity by restoring liver function to normal (95%effect). The cell toxicity indicators shown in the said FIG. 1 is theleakage of lactate dehydrogenase (LDH) from intact cells after toxinchallenge and its reversal by extracts.

FIG. 2 shows leakage of glutamate pyruvate transaminase wherein saidplant extracts in combination attenuates Rifampicin+isoniazid inducedhepatotoxicity by restoring liver function to normal (96%). The celltoxicity indicators shown in FIG. 2 is the leakage of glutamate pyruvatetransaminase (GPT) from intact cells after toxin challenge and itsreversal by the extracts in combination. (Model: primary monolayercultures of liver cells).

FIG. 3 shows leakage of serum glutamate pyruvate transaminase (GPT)after toxin challenge and its reversal by Emblica officinalis fractionwherein Emblica officinalis reverses Rifampicin+isoniazid+pyrazinamideinduced hepatotoxicity and restores the liver function to normal (96%).The cell toxicity indicators shown in FIG. 3 is the leakage of serumglutamate pyruvate transaminase (GPT) after toxin challenge and itsreversal by Emblica officinalis fraction.

FIG. 4 shows protection against cell leakage wherein, Emblicaofficinalis fractions in combination prevents Rifampicin+isoniazidinduced toxicity. FIG. 4 shows that fractions provide 96% protectionagainst cell leakage as measured by serum GPT levels and increasescellular defense. 75% increase in glutathione levels (liver) isobserved.

FIG. 5 shows Emblica officinalis extract reversesRifampicin+isoniazid+pyrazinamide induced toxicity. FIG. 5 shows 94%protection against rise in lipid Peroxidation (LPO, liver) and 96%decrease of serum Bilirubin in response to treatment with extract of thepresent invention.

FIG. 6 shows a flow chart for the preparation of extract from fruitEmblica Officinalis.

EXAMPLES

The invention of instant Application is further illustrated by thefollowing examples, which should not, however be construed to limit thescope of the invention. The following examples are not intended to belimiting in any way, but demonstrate some of the preferred embodimentsof the present invention. Any person skilled in the art can design morecombinations useful against drug-induced toxicity, which may beconsidered as part of the present invention.

Example 1

Table 1 shows that the plant products are effective against paracetamolhepatotoxicity which is mainly dependent against bioactivationmechanisms mediated by CYP 450.% protection is shown as combined effectrelease of LDH and GPT in serum. TABLE 1 Effect of plant productsagainst hepatotoxicity produced by paracetamol Treatment paracetamolToxicity 93% % protection as measured by combined effect against LDH andGPT leakage Extract 97% protection Fraction 96% protection

Example 2

Table 2 shows that the preparations of the present invention are noteffective against liver toxicity produced by agents where the toxicityis primarily not dependent on bioactivation by CYP 450. TABLE 2 Effectof plant products against hepatotoxicity produced by galactosamine.Treatment galactosamine Toxicity 93% % protection as measured bycombined effect against LDH and GPT leakage Extract  3% Fraction  7%

Example 3

Plant extract attenuates Rifampicin induced hepatotoxicity by restoringliver function to normal (95% effect). The cell toxicity indicatorsshown in FIG. 1 is the leakage of lactate dehydrogenase (LDH) fromintact cells after toxin challenge and its reversal by extracts. (Model;primary monolayer cultures of liver cells)

Example 4

Plant extracts in combination attenuates Rifampicin+isoniazid inducedhepatotoxicity by restoring liver function to normal (96%). The celltoxicity indicators shown in FIG. 2 is the leakage of glutamate pyruvatetransaminase (GPT) from intact cells after toxin challenge and itsreversal by the extracts in combination. (Model: primary monolayercultures of liver cells).

Example 5

Emblica officinalis reverses Rifampicin+isoniazid+pyrazinamide inducedhepatotoxicity and restores the liver function to normal (96%). The celltoxicity indicators shown in FIG. 3 is the leakage of serum glutamatepyruvate transaminase (GPT) after toxin challenge and its reversal byEmblica officinalis fraction.

Example 7

Emblica officinalis fractions in combination preventsRifampicin+isoniazid induced toxicity. FIG. 4 shows that fractionsprovide 96% protection against cell leakage as measured by serum GPTlevels and increases cellular defense. 75% increase in glutathionelevels (liver) is observed.

Example 7

Emblica officinalis extract reverses Rifampicin+isoniazid+pyrazinam−ideinduced toxicity. FIG. 5 shows 94% protection against rise in lipidPeroxidation (LPO, liver) and 96% decrease of serum Bilirubin inresponse to treatment with extract of the present invention.

1. An orally-administered composition useful for treatment of CYP 450bio-activated hepatotoxicity induced by at least one anti-TB drugselected from the group consisting of rifampicin, pyrazinamide, andisoniazid so as to restore liver function to normal, said compositioncomprising an extract from Emblica officinalis and pharmaceuticallyacceptable additives, wherein the extract comprises from 8.5% to 15% byweight of tannin, said extract and said additives being in the ratio of1:1-1:10, the additives being selected from a group of nutrientscomprising proteins, carbohydrates, sugar, talc, magnesium stearate,cellulose, calcium carbonate, and starch-gelatin paste, and/orpharmaceutically acceptable carrier, excipient, diluent, or solvent,wherein said composition treats CYP 450 bio-activated hepatotoxicityinduced by at least one anti-TB drug selected from the group consistingof rifampicin, pyrazinamide, and isoniazid so as to restore liverfunction to normal, but is not effective against hepatotoxicity which isindependent of bio-activation by CYP
 450. 2-4. (canceled)
 5. Acomposition as claimed in claim 1, wherein said additives have no effecton the hepatocurative effect of the said extract.
 6. A composition asclaimed in claim 1, wherein said extract is prepared in a solventselected from a group comprising aqueous, aqueous-ethanolic, ethanolic,ketonic, ethereal, halogenated solvents.
 7. (canceled)
 8. A compositionas claimed in claim 1, wherein, said composition for the oral route isin form of capsule, tablet, syrup, concentrate, powder, granule,aerosol, and/or beads.
 9. A method of preparing an orally-administeredcomposition useful for treatment of CYP 450 bio-activated hepatotoxicityinduced by at least one anti-TB drug selected from the group consistingof rifampicin, pyrazinamide, and isoniazid so as to restore liverfunction to normal, said composition comprising an extract from Emblicaofficinalis and pharmaceutically acceptable additives, wherein theextract comprises from 8.5% to 15% by weight of tannin, said extract andsaid additives being in the ratio of 1:1-1:10, the additives beingselected from a group of nutrients comprising proteins, carbohydrates,sugar, talc, magnesium stearate, cellulose, calcium carbonate, andstarch-gelatin paste, and/or pharmaceutically acceptable carrier,excipient, diluent, or solvent, wherein said composition treats CYP 450bio-activated hepatotoxicity induced by at least one anti-TB drugselected from the group consisting of rifampicin, pyrazinamide, andisoniazid so as to restore liver function to normal, but is noteffective against hepatotoxicity which is independent of bio-activationby CYP 450, said method comprising incubating said Emblica officinaliswith a polar solvent at room temperature for about 15 to 25 hours so asto obtain the extract, the extract comprising from 8.5% to 15% by weightof tannin, and optionally adding pharmaceutically acceptable additives,wherein the polar solvent consists of (i) water, (ii) water and ethanolor (iii) ethanol.
 10. (canceled)
 11. A method as claimed in claim 9,wherein extract is from fresh and/or semi dried fruits of EmblicaOfficinalis. 12-13. (canceled)
 14. A method as claimed in claim 9,wherein said composition is in form of capsule, tablet, syrup,concentrate, powder, granule, aerosol, and/or beads. 15-38. (canceled)